Citation: HONG Xiao-Yu, WANG Hao, XU Jin-Ling, LI Shui-Ming, WANG Yong. Application of High Resolution Time-of-Flight Mass Spectroscopy in Relative Quantitative Analysis in Proteomics[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(3): 403-408. doi: 10.11895/j.issn.0253-3820.150751 shu

Application of High Resolution Time-of-Flight Mass Spectroscopy in Relative Quantitative Analysis in Proteomics

1.
¹ College of Life Science, Shenzhen Key Laboratory of Microorganism, Shenzhen University, Shenzhen 518060, China;
2.
² College of Life Science, Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen University, Shenzhen 518060, China

Corresponding author: WANG Yong,
Received Date: 22 September 2015
Available Online: 12 December 2015
Fund Project: 本文系深圳市科技项目(Nos.JSGG20140703163838793,20150006)资助 (Nos.JSGG20140703163838793,20150006)

By using the high resolution mass spectrometer TripleTOF 5600, three kinds of standard proteins including bovine serum albumin (BSA), ovalbumin (OVA) and lysozyme C(LYZC) were analyzed, and the correlationship between the ion intensity of mass spectrometry and the relative content of protein sample was investigated. The protein samples were digested by trypsion and diluted to 1-1024 fmol in 7 μL. The ion counts per second (cps) were used to stand for the amounts of proteins and peptides. Then the correlation between sum of ion intensity (cps) of all the peptides, number of peptides detected and the amount of proteins was investigated. By comparing the change of values of the same sample in three parallel experiments, a linear relationship between these indexes and the amount of proteins within 1-1024 fmol was found when the cps was more than 1000. Usually, the maximal ion intensity was no more than 1.5 times of the minimum value for same peptide in triplicate experiments, which suggested that the 3 times or more change of ion intensity was the minimum threshold to determine the differences of proteins amounts in different samples. This study provides a relative quantitative analysis method using qualitative data of high resolution and high scan speed mass spectrometry, which can quickly and easily provide reference for biological and medical research.
- Quantitative proteomics,
- High resolution time-of-flight mass spectrometry,
- Counts per second,
- Signal intensity,
- Label-free quantification
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沈阳化工大学材料科学与工程学院沈阳 110142